D. Moigradean, et all.

Journal of Agroalimentary Processes and Technologies,

Volume XII, No. 2 (2006), 385-390
Full Paper - Food Control Section

ANTIOXIDANT CAPACITY EVALUATION IN RELATION

WITH POLYPHENOLOS AND ASCORBIC ACID CONTENT
FOR SOME NATURAL JUICES
Diana Moigrădean, Mariana-Atena Poiană, I. Gergen, Diana Dogaru
Banat′s University of Agricultural Sciences and Veterinary Medicine, Faculty of
Food Processing Technology, Calea Aradului no.119, RO-300645, Timisoara,
Romania

Abstracts
Twelve different commercial natural juices of fruits (six nectar and
six clear juices) from Timisoara market were analyzed for some
chemical characteristics (total acidity, soluble substance, total extract-
determined on the basis of standardized method) and antioxidant
activities (using FRAP method), total polyphenols amount (by Folin-
Ciocalteu method) and ascorbic acid content (using 2,6-
dichlorophenol indophenol method). The antioxidant capacity varied
between 1.5-3 mM Fe2+/L for clear natural juices and between 4-6.5
mM Fe2+/L for fruit nectar. The polyphenols content it was situated
between 0.8-1.1 mM acid gallic/L for clear natural juices and
between 2-5 mM acid gallic/L for fruit nectar. The values obtained for
ascorbic acid content was situated between 0.5-2.2 mM/L, different for
clear juices and nectar.
Key words: antioxidant capacity, polyphenols, ascorbic acid, fruit
nectar, clear natural juice
Introduction
The natural juices are of great interest because of their potent
antioxidant capacity and possible protective effects on human health.
Flavonoids in fruits, vegetables and juices appear to play a significant
role in cancer and heart disease health benefits, and proanthocyanidins
account for a significant portion of the total flavonoids ingested by
peoples. Antioxidants have been shown to prevent the oxidation of
cholesterol, especially the bad cholesterol, low-density lipoprotein
(LDL). High cholesterol is, of course, a scientifically documented risk
factor for heart disease. Recently LDL oxidation has been
hypothesized as the beginning of the atherosclerosis process that starts

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 Antioxidant Capacity Evaluation in Relation with Polyphenolos and Ascorbic Acid
Content for Some Natural Juices

in childhood. In short, your pipes begin to rust at an early age

(Dragsted, 2003; Facino, 1999; Fitzpatrick, 2002).
In according to the many authors (Simonetti, 2002; Tosun, 2003),
people drink fruit juice for many reasons: orange juice, for example, is
an excellent source of Vitamin C, folate and potassium nutrients, many
people may consume in sufficient quantity in their diets otherwise. But
for people looking for dietary approaches to increase consumption of
natural antioxidants that may help reduce the risk of heart disease,
purple grape juice appears unique in its potential ability to provide a
beneficial effect to a variety of cardiovascular functions. By drinking
this juice, you are going to give yourself the antioxidant profile you
need to protect your arteries, your brain cells, and literally every part
of your body from oxidation. You will also maximize and optimize
your health and energy levels with something that actually tastes
delicious.
The purpose of this paper was to determine the total antioxidant
capacity and to found some correlations between total polyphenos
amount, ascorbic acid and antioxidant capacity for natural juices from
Romanian market. Fruit polyphenols include a wide range of
compounds with antioxidant activity, that is: anthocyanins, flavonols,
flavan-3-ols, hydroxycinnamates (Harmanescu, 2005).
The measurement of total antioxidant capacity (TAC) can be
making using different methods (photometric, chemiluminescence’s
and spectrophotometric methods). On the basis of the mentioned data
in literature and on the experience of the authors regarding the
determination of antioxidant capacity, we chose the modified method
FRAP. This method it was based of the ability of antioxidant
substances present in analyzed juices to reduce the ferric ions (Fe3+)
from tripyridyltriazine complex to the ferrous ions (Fe2+). The ferrous
tripyridyltriazine complex has an intensive blue color and can be
monitored at 593 nm (Benzie & Strain, 1996).
This method is properly in the case of total antioxidant capacity
determination for different natural products (Tosun, 2003; Gergen,
2004, Gergen, 2005).
For polyphenols determination (expressed such as mM/L acid
galic) will be use the spectroscopic method with the Folin Ciocalteu
reactiv (Folin, 1927; Gergen, 2004; Gergen, 2005).

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 D. Moigradean, et all. Journal of Agroalimentary Processes and Technologies,
Volume XII, No. 2 (2006), 385-390

Experimental
Reagent and equipment: All chemicals and reagents were
analytical grade or purest quality purchased from Merck, Fluka,
Sigma. Was used distilled water. Absorption determination for FRAP
and total phenol content was made using Spectrophotometer Specord
205 by Analitik Jena.
Nectar and clear drinks: In the present research, a total six fruit
clear natural juices (red orange, orange, tutti frutti,
strawberry&banana, grapefruit, pine-apple) and six nectars (apricot,
pear, orange, peach, cherry, tutti frutti) were investigated. These juices
were processed of S.C. EUROPEAN DRINKS S.A., Bihor, Romania
(FRUTTIA mark) and purchased from Timisoara supermarkets.
Determination of Total Antioxidant Capacity (Adaptation of FRAP
method): Reagents: acetate buffer, 300 mM/L, pH = 3.6 (3.1 g sodium
acetate 3H2O and 16 mL conc. Acetic acid per 1L off buffer solution);
TPTZ (2,4,6-tripyridyl-s-triazine) solution 10 mM/L (0.31 g TPTZ in
100 mL HCl). Prepared freshly always utilization; FeCl3 solution
20mM/L (0.54 g FeCl3·6H2O in 100 mL distilled water). Prepared
freshly always utilization; FRAP working solution (25 mL acetate
buffer, 2.5 mL TPTZ solution, 2.5 mL FeCl3 solution). Prepared
freshly always utilization; Standard solution - Mohr salt 1mM/L.
Aqueous solution of known Fe concentration was used for
calibration, in a range of 0.1-0.8 mM/L. For the preparation of
calibration curve 0.5 mL aliquot of 0.1, 0.2, 0.4, 0.6, 0.8 µM Fe2+/mL
aqueous as Mohr salt solution were mixed with 2.5 mL FRAP working
solution. FRAP reagent was used as blank. One mL from diluted 1/10
juices and nectars was mixed with the same reagents as described
above, and after 10 min. absorption was read after at λ= 593 nm.
The Total antioxidant capacity in fruit juices and fruit nectar in Fe
(II) equivalents was calculated. Correlation coefficient (r2) for
calibration curve was 0.998.
Determination of phenolic compounds: The content of total
polyphenolic compounds was determined by Folin-Ciocalteu method.
Reagents: Folin-Ciocalteu′s phenol reagent solution 1:10; Na2CO3
solution 7.5%; Standard solution - Gallic acid 10mM/L. For the
preparation of calibration curve 0.5 mL aliquot of 0.2, 0.3, 0.4, 0.8 and
1.2 µM/mL aqueous gallic acid solution were mixed with 2.5 mL

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 Antioxidant Capacity Evaluation in Relation with Polyphenolos and Ascorbic Acid
Content for Some Natural Juices

Folin-Ciocalteu reagent and 2.0 mL sodium carbonate. One mL from

diluted 1/10 juices and nectars was mixed with the same reagents as
described above, and absorption was read after 2 h at λ= 750 nm. Total
content of polyphenols in fruit juices and fruit nectar in gallic acid
equivalents (GAE) was calculated. Correlation coefficient (r2) for
calibration curve was 0.995.
Determination of ascorbic acid: Ascorbic acid content was
estimated titrimetrically with 2,6-dichlorphenolindophenol natrium dye
reactant. 10 mL of clear natural juice (diluted 1/10) or nectar (diluted
1/20) with 10 mL distilled water and 1 mL HCl 1N it was titrated with
2,6-dichlorphenolindophenol natrium to pink color. On the basis of
2,6-dichlorphenolindophenol natrium volum it was determined the
ascorbic acid amount. [Szeto, 2002; Gergen, 2004]
Results and Discussions
The commune chemical characteristics (total acidity, soluble
substance, total extract) of natural juices are presented in Table 1.
Table 1. Total acidity, soluble substance and total extract of some clear
juices
Nr. Natural juice Total acidity Soluble Extract
crt. ( g malic acid /100 mL) substance (°Bx) (g/L)
Clear juice
1. Red Orange 0.43 11.2 11.5
2. Orange 0.36 10.2 10.4
3. Tutti Frutti 0.45 11.2 11.5
4. Strawberry& 0.17 11.4 11.7
banana
5. Grapefruit 0.25 10.2 10.5
6. pine-apple 0.25 10.6 11.0
Nectars
1 Apricot 0.88 15.4 18.2
2 Pear 0.47 15.8 17.4
3 Orange 0.86 13.0 13.8
4 Peach 0.70 15.9 19.7
5 Cherry 0.60 14.1 16.4
6 Tutti Frutti 0.61 13.1 14.9

The values for characteristics showed in table 1 are in according to

the natural juices type (clear juices or nectars). The fruit percent in

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 D. Moigradean, et all. Journal of Agroalimentary Processes and Technologies,
Volume XII, No. 2 (2006), 385-390

clear juices is min. 5-10% while in nectar min. 35-50%. This fruit
percent is proceeded to concentrate from natural fruit juice.
The total antioxidant capacity–TAC by FRAP method, Ascorbic
Acid and polyphenols are presented in Table 2 for soft drinks and in
Table 3 for analyzed nectars.
Table 2. Antioxidant capacity, polyphenols and Vitamin C content of some
clear drinks
Antioxidant
Poliphenols Ascorbic acid
Clear juice capacity
(mM acid gallic/L) (mM/L)
(mM Fe2+/L)
Red Orange 2.57 1.13 0.88
Orange 2.48 0.91 0.96
Tutti Frutti 2.87 1.12 1.04
Strawberry & banana 1.40 0.58 0.48
Grapefruit 2.02 0.83 0.72
pine-apple 2.36 1.00 0.96
Table 3. Antioxidant capacity, polyphenols and Vitamin C content of some
nectars
Antioxidant capacity Poliphenols Ascorbic acid
Nectars
(mM Fe2+/L) (mM acid gallic/L) (mM/L)
Apricot 4.64 3.9 1.28
Pear 5.27 3.56 1.60
Orange 4.90 2.26 2.24
Peach 4.56 3.86 1.76
Cherry 6.52 5.20 1.28
Tutti Frutti 4.30 3.18 2.08

The nectars present TAC more then clear juices. The same
variation was observed for total polyphenols and ascorbic acid. For
nectar, the highest polyphenols amount was found in cherry nectar and
the smaller in orange. For clear juices, the highest polyphenol content
is in tutti frutti juice and the smaller value was identified in strawberry
&banana juices. For nectar, the ascorbic acid content is highest in
orange and minim in apricot and cherry. For clear juices, the smaller
values were identified in strawberry&banana juice and the maxim
values in tutti frutti. We explained the different values for these
characteristics on the basis of applied processing technology (for clear
juices the phenolic compound were removed by clearing operation).

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 Antioxidant Capacity Evaluation in Relation with Polyphenolos and Ascorbic Acid
Content for Some Natural Juices

Conclusions
The fruit nectars were presented more total antioxidant activity,
polyphenols and ascorbic acid content than clear natural juices. For
clear natural juices, TAC is very good correlated with the sum of
ascorbic acid and polyphenols content. For analyzed nectars, the
correlation is not was observed. The results of this research show that,
the natural juices have an excellent source of natural antioxidants
being consumed with pleasure because are more readily assimilated
than fruit tissues.
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